ObjectiveTo investigate the effects of overexpression of alpha/beta hydrolase domain-containing protein 5 (ABHD5) on the invasion and migration of human colon cancer cell line HCT116 and the pathway of adenosine monophosphate-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR).MethodsThe expression of ABHD5 in colon cancer tissues and its relationship with clinicopathological features was analyzed by UALCAN database. HCT116 cells were cultured in vitro and transfected with ABHD5 recombinant plasmid, then they were divided into control group, negative transfection group and ABHD5 transfection group. Real time quantitative PCR (qRT-PCR) was used to detect the expression of ABHD5 mRNA in HCT116 cells. The proliferation of HCT116 cells was detected by CCK-8 method. Transwell assay was used to detect the invasion and migration of HCT116 cells. The expression of matrix metalloprotein 9 (MMP-9), E-cadherin, Snail, and AMPK/mTOR pathway proteins p-AMPK, AMPK, p-mTOR and mTOR were detected by Western blot.ResultsThe results of the UALCAN showed that compared with normal colon tissues, the expression of ABHD5 mRNA in colon cancer tissues was decreased (P<0.05), and which in the adenocarcinoma and the N1 stage was lower than that of the mucinous adenocarcinoma (P<0.05) and N0 stage (P<0.05), respectively. Compared with the control group and the negative transfection group, the expression of ABHD5 mRNA in the ABHD5 transfection group was increased (P<0.05), the proliferation inhibition rate of HCT116 cells in the ABHD5 transfection group was increased (P<0.05), the numbers of migration and invasion cells in the ABHD5 transfection group were decreased (P<0.05), the expressions of MMP-9, Snail, p-mTOR and mTOR were reduced, and the expressions of E-cadherin, p-AMPK and AMPK were increased (P<0.05).ConclusionsThe overexpression of ABHD5 can inhibit the invasion and migration of colon cancer HCT116 cells, activate AMPK, and inhibit the expression of mTOR. It suggests that ABHD5 may play a role in inhibiting colon cancer by affecting AMPK/mTOR pathway.
ObjectiveTo detect the expressions of takeda G protein-coupled receptor 5 (TGR5) and mortalin protein 75 in the tissues of patients with intrahepatic cholangiocarcinoma (ICC), and to explore their relationship with prognosis.MethodsA total of 94 ICC patients who were admitted to Anyang District Hospital and received surgical treatment from March 2015 to March 2018 were selected as the research objects. The expressions of TGR5 and mortalin protein 75 in ICC cancer tissues and adjacent tissues were detected by immunohistochemistry and Western blot (WB). The relationship between the expressions of TGR5 and mortalin protein 75 in ICC cancer tissues and clinicopathological parameters and prognosis was analyzed. Multivariate Cox proportional hazards regression was used to analyze the risk factors of poor prognosis in patients with ICC. ROC curve was used to analyze the diagnostic value of TGR5 and mortalin protein 75 for poor prognosis in patients with ICC.ResultsImmunohistochemical results showed that the positive expression rates of TGR5 and mortalin protein 75 in cancer tissues were significantly higher than those in adjacenttissues (P<0.05). WB results showed that the protein expression levels of TGR5 and mortalin protein 75 in cancer tissues were significantly higher than those in adjacent tissues (P<0.05). The expression of TGR5 protein in cancer tissues of ICC patients was correlated with gender, tumor diameter, degree of differentiation, TNM staging, satellite focus, and liver cirrhosis (P<0.05). The expression of mortalin protein 75 was correlated with tumor diameter, TNM staging, nerve involvement, satellite focus, and liver cirrhosis (P<0.05). There were significant differences in gender, tumor diameter, TNM staging, microvascular invasion, satellite focus, liver cirrhosis, and the expressions of TGR5 and mortalin protein 75 between the poor prognosis group and the good prognosis group (P<0.05). The cumulative 3-year overall survival rate of TGR5 positive patients (32.00%) was significantly lower than that of TGR5 negative patients (63.16%), χ2=6.228, P=0.013; the cumulative 3-year overall survival rate of mortalin protein 75 positive patients (32.91%) was significantly lower than that of mortalin protein 75 negative patients (66.67%), χ2=6.079, P=0.014. Multivariate Cox proportional hazards regression analysis showed that the positive expression of TGR5 and mortalin protein 75, TNM Ⅲ+Ⅳphase, satellite focus, and cirrhosis were risk factors for poor prognosis in ICC patients (P<0.05). ROC results showed that when the expression level of TGR5 was 0.932 as the cut-off value, its AUC in the diagnosis of poor prognosis of ICC patients was 0.783, the sensitivity was 72.4%, the specificity was 72.2%; when the expression level of mortalin protein 75 was 0.756 as the cut-off value, its AUC in the diagnosis of poor prognosis of ICC patients was 0.805, the sensitivity was 84.4%, the specificity was 63.9%; the AUC of combined diagnosis of TGR5 and mortalin protein 75 was 0.884, the sensitivity was 79.3%, the specificity was 83.3%.ConclusionsThe high expressions of TGR5 and mortalin protein 75 in cancer tissues of ICC patients are associated with poor prognosis, and they are risk factors for poor prognosis. The combined detection of TGR5 and mortalin protein 75 has a certain value in predicting poor prognosis, and can be used as potential biological indicators.